Anti-GFP antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (AB6556)

This image shows a single primary hippocampal neuron from a primary culture overexpressing GFP stained with ab6556 at a dilution of 1/2000. This picture was kindly supplied as part of the review submitted by one of our customers.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (AB6556)

This image shows IF using GFP-expressing glial cells (green) transplanted into lesioned rat spinal cord. This was detected using ab6556 anti-GFP antibody and a FITC conjugated secondary antibody. Axons are labelled red by an antibody to neurofilament-200 and a rhodamine secondary. ab6556 reveals the morphology of the transplanted cells to such an extent that their close interactions with axons are obvious. The top picture shows an optical section from a confocal microscope scan showing how a GFP cell wraps around a branched axon travelling longitudinally. The bottom picture consists of an optical section from another confocal scan showing a GFP cell enveloping an axon in the transverse plane. Review by Andrew Toft submitted 19 May 2004.

• EM

Unknown

Electron Microscopy - Anti-GFP antibody (AB6556)

Specific labeling of a Trk-GFP fusion protein being synthesized on ER in sympathetic neurons infected with an adenovirus carrying the construct. The gold is associated with the ER membranes. This was done using a 1/5000 dilution of affinity purified antibody (ab6556). The tissue section was fixed and embedded using durcupan resin.

• IHC - Wmt

CiteAb

IHC - Wholemount - Anti-GFP antibody (AB6556)

GFP immunohistochemistry-immunofluorescence using Anti-GFP antibody ab6556. Publication image and figure legend from Wilm, T. P., Tanton, H., et al. 2021, Sci Rep, PubMed 34354169.

Whole mount and histological analysis of adult kidney after lineage tracing of Wt1-expressing cells. Adult mice with either the Wt1CreERT2/+; Rosa26LacZ/LacZ or the Wt1CreERT2/+; Rosa26mTmG/mTmG reporter system were analysed 2-4 weeks after Tamoxifen administration. (A,B) In kidney whole mounts (sagittal halves), labelled cells expressing LacZ or GFP were found in the glomeruli. (C) Eosin-counterstained sagittal paraffin sections showing LacZ-expression in the glomeruli of the kidney (open arrowheads pointing to glomeruli showing weaker XGal staining due to reduced penetration of staining reagents into tissue). (C',C") LacZ-expressing cells are also detected in the parietal epithelial layer of the Bowman Capsule (filled arrowheads). (D) Immunofluorescence on frozen sagittal kidney sections revealed GFP-expressing cells in the glomeruli. (E) Immunofluorescence for Wt1 and GFP in higher magnification of the two glomeruli outlined in the box in (D). (E'-E"") Immunofluorescence for Wt1 and GFP of the left glomerulus shown in (E). GFP-labelled cells co-expressed Wt1 (E', Wt1 and DAPI; E", GFP and DAPI; E", Wt1 and GFP; E"", Wt1, GFP, DAPI). (F,G) In rare cases, LacZ- or GFP-expressing cells were found in tubular structures reaching into the renal medulla (filled arrowheads). The data shown in (A-E) are consistent with analyses performed in n = 5 animals. Scale bars, 2 mm (A,B), 300 µm (C,D), 100 µm (C',C",E), 50 µm (E'-E""), 700 µm (F,G).

• IHC

CiteAb

Immunohistochemistry - Anti-GFP antibody (AB6556)

GFP Immunohistochemistry using Anti-GFP antibody ab6556. Schipper, K., Seinstra, D., et al., 2019, Nat Commun, PubMed 31444332.

E-cadherin loss drives cell extrusion towards the basal lamina. a Schematic overview of engineered alleles in Wcre;Cdh1F/F;mTmG mice. b, c Examination of GFP-positive Wcre activity in mammary glands of 6-week-old Wcre;mTmG female mice by immunofluorescence (IF) analysis (n = 6). b IF staining of GFP was examined in cytokeratin-8 (CK8)-positive luminal mouse mammary epithelial cells (MMECs) and c cytokeratin-14 (CK14)-positive myoepithelial cells. Scale bar, 10 µm. d Identification of E-cadherin inactivated MMECs by IF staining of E-cadherin, GFP, CK14, and CK8 in 6-week-old Wcre;Cdh1F/F;mTmG female mice (n = 6). Arrows indicate events of cell extrusion upon inactivation of E-cadherin. Scale bare, 10 µm. e Immunohistochemical detection of GFP-positive E-cadherin inactivated MMECs in mammary gland sections of 3-, 4-, and 6-month-old Wcre;Cdh1F/F;mTmG female mice and age-matched WCre;mTmG control mice (n = 3). Scale bar, 20 µm. f Examination of E-cadherin expression in mammary gland sections of 3-month-old Wcre;Cdh1F/F;mTmG female mice and age-matched Wcre;mTmG control mice by IF analysis of GFP, E-cadherin, CK14, and Hoechst. Asterisk indicates area of zoom. Scale bar, 50 µm. g Quantification of the amount of extruded GFP-positive cells in 3-month-old Wcre;Cdh1F/F;mTmG (n = 3) female mice and age-matched Wcre;mTmG (n = 3) control mice. Data are of nine images per group. h Quantification of the average size of extruded GFP-positive cell clusters in the mammary glands of Wcre;Cdh1F/F;mTmG mice at the ages of 3, 5, and 12 months. Data are of three mice per time point and 10 images per mouse. All data are depicted as mean ± standard deviation. All p values were calculated using an unpaired two tailed t-test. Source data are provided as a Source Data file

• Flow Cyt

CiteAb

Flow Cytometry - Anti-GFP antibody (AB6556)

GFP flow cytometry using Anti-GFP antibody ab6556. Publication image and figure legend from Nakagawa, A., Naito, A. T., et al., 2016, Sci Rep, PubMed : 27146149.

Inducible activation of Wnt/β-catenin signaling in arterial ECs.(a) Flow cytometric analysis of ECs. ECs were collected from the heart of Bmx-CreERT2 mice (Ctrl) or from Bmx-CreERT2 crossed with CAG-CAT-EGFP mice (Bmx/EGFP) 1 week after the TAM treatment. (b) Immunofluorescent staining of cardiac tissue for GFP (red), CD31 (green), and TO-PRO-3 (blue). Scale bars : 20 μm. (c, d, e) Genotyping PCR (c), western blot (d) and quantitative RT-PCR analysis (e) of cardiac ECs isolated from Ctrl (Bmx-CreERT2 with Ctnnb1+/+) mice (Ctrl ECs) and Bmx/CA mice (Bmx/CA ECs). (c) Floxed allelle of β-catenin (=500 bp) was detected in Bmx/CA ECs but not in Ctrl ECs. (d) β-catenin protein lacking exon3 (=75 kDa) was detected in Bmx/CA ECs but not in Ctrl ECs. (e) Expression levels of Wnt/β-catenin signaling target genes (Axin2 and Lef1) were higher in Bmx/CA ECs compared with Ctrl ECs. **p < < 0.01.

• WB

CiteAb

Western blot - Anti-GFP antibody (AB6556)

GFP western blotting using Anti-GFP antibody ab6556. Publication image and figure legend from Rivero-Ríos, P., Romo-Lozano, M., et al., 2020, Cells PubMed 32709066.

Deficits in EGF trafficking due to knockdown of RAB8A or RAB10 are rescued upon RAB29 expression. (A) HeLa cells were either transfected with ctrl-siRNA or RAB10-siRNA, and cotransfected with GFP-tagged RAB29 constructs as indicated, and the amount of surface-bound fluorescent EGF quantified. N = 3 independent experiments; * p < < 0.05. (B) Cells were transfected with ctrl-siRNA or RAB10-siRNA, and cotransfected with GFP-tagged RAB29 constructs as indicated, and internalized fluorescent EGF was quantified at 10 min (left) and 30 min (right). N = 3 independent experiments; *** p < < 0.005; **** p < < 0.001. (C) Cells were either transfected with ctrl-siRNA or RAB10-siRNA, and transfected with GFP-tagged RAB29 constructs as indicated, and cell extracts (30 μg) were analyzed by Western blotting for GFP-RAB29 levels, endogenous RAB10 levels, and GAPDH as loading control. (D) HeLa cells were either transfected with ctrl-siRNA or RAB8A-siRNA, and cotransfected with GFP-tagged RAB29 constructs as indicated, and the amount of surface-bound fluorescent EGF was quantified. N = 3 independent experiments; * p < < 0.05. (E) Cells were transfected with ctrl-siRNA or RAB8A-siRNA, and cotransfected with GFP-tagged RAB29 constructs as indicated, and internalized fluorescent EGF was quantified at 10 min (left) and 30 min (right). N = 3 independent experiments; * p < < 0.05; ** p < < 0.01; *** p < < 0.005. (F) Cells were either transfected with ctrl-siRNA or RAB8A-siRNA, and transfected with GFP-tagged RAB29 constructs as indicated, and cell extracts (30 μg) were analyzed by Western blotting for GFP-RAB29 levels, endogenous RAB8A levels, and tubulin as loading control.

false